title: Menthol derivative WS-12 selectively activates transient receptor potential melastatin-8 (TRPM8) ion channels.
abstract: Transient receptor potential melastatin-8 (TRPM8), a cationic ion channel is involved in detection of normal cooling-sensation in mammals. TRPM8 activation by cooling or chemical agonists have been shown to produce profound, mechanistically novel analgesia in chronic pain states such as neuropathic pain in rodents. Known TRPM8 agonists such as menthol and icilin have a relatively low potency and cross-activate nociceptors like TRPA1; thus bearing a limited therapeutic usefulness. For that reason, characterising ligands, which selectively activate TRPM8, presents a clinical need. Using Xenopus laevis oocytes as expression system, we evaluated WS-12, a menthol derivative, for its potential interaction with all six thermo-sensitive TRP ion channels. Oocytes were injected with cRNA of gene of interest and incubated for 3-5 days (at 16 degrees C) before testing for functional characterisation of the recombinant ion channels. Oocytes were superfused with the test and standard substances respectively. Responses were measured by two-electrode voltage clamp technique and the amplitudes of evoked currents were compared with baseline values. WS-12 robustly activated TRPM8 in low micromolar concentrations (EC50 12+/-5 microM) thereby displaying a higher potency and efficacy compared to menthol (EC50 196+/-22 microM). Any of the other described thermo-sensitive TRP ion channel including TRPV1, TRPV2, TRPV3, TRPV4 and TRPA1 were not activated at a concentration (1 mM) optimally effective for TRPM8 responses; a characteristic which is in sharp contrast to menthol as it activates TRPA1 and TRPV3 in addition to TRPM8. Unlike icilin (75% reduction; p<0.001, n=6), WS-12 does not induce tachyphylaxis (4+/-2.3% increase in responses; p<0.08, n=6) of TRPM8 mediated currents to repeated exposure of 1 mM doses. In addition, acidosis or variations in extracellular calcium have no influence on potency/efficacy of WS-12 for TRPM8. The selectivity profile of WS-12, its several-fold higher potency and around two-fold increase in efficacy compared to menthol warrants its potential utility for therapy in chronic neuropathic pain states and as a diagnostic probe in prostate cancer.
authors.affiliation: Department of Cell Physiology, Faculty of Biology & Biotechnology, Ruhr-University-Bochum, University Street 150, Bochum 44801, Germany.
authors.forename: Sherkheli
authors.initials: S
authors.lastname: Ma
authors.forename: Gisselmann
authors.initials: G
authors.lastname: G
authors.forename: Vogt-Eisele
authors.initials: VE
authors.lastname: Ak
authors.forename: Doerner
authors.initials: D
authors.lastname: Jf
authors.forename: Hatt
authors.initials: H
authors.lastname: H
mesh_terms: D000700:Analgesics / Q000494:pharmacology*; D000813:Anilides / Q000494:pharmacology*; D000818:Animals; D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels; D004305:Dose-Response Relationship, Drug; D018014:Gene Transfer Techniques; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008564:Membrane Potentials; D008610:Menthol / Q000031:analogs & derivatives* / Q000494:pharmacology; D051379:Mice; D009419:Nerve Tissue Proteins / Q000819:agonists; D011744:Pyrimidinones / Q000494:pharmacology; D051381:Rats; D000074025:TRPA1 Cation Channel; D050053:TRPM Cation Channels / Q000819:agonists* / Q000378:metabolism; D050916:TRPV Cation Channels / Q000819:agonists; D013618:Tachyphylaxis; D050051:Transient Receptor Potential Channels / Q000819:agonists; D014982:Xenopus laevis
chemical_list: D000700:Analgesics; D000813:Anilides; D015220:Calcium Channels; C533807:N-(4-methoxyphenyl)-4-menthone-3-carboxamide; D009419:Nerve Tissue Proteins; D011744:Pyrimidinones; D000074025:TRPA1 Cation Channel; C473350:TRPA1 protein, human; D050053:TRPM Cation Channels; C492297:TRPM8 protein, mouse; D050916:TRPV Cation Channels; D050051:Transient Receptor Potential Channels; C486760:Trpv3 protein, mouse; D008610:Menthol; C490483:icilin; D002118:Calcium